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• W2006104692 abstract "Early detection and genotyping of HCV infection is of highest importance for the management of the disease (1). Until recently, the HCV detection and quantification in our laboratory was done by AMPLICOR HCV tests while genotyping was performed with combined AMPLICOR HCV v2.0 test/ASO hybridization method with probes for genotypes 1–4 (2). We were looking to reduce the cost and average time necessary for reaching a diagnosis by switching to semi-automated techniques allowing an accurate, sensitive, cost-effective and reproducible assays for detection, quantification and genotyping of all the relevant HCV variants in our country simultaneously. The combined methodology for HCV diagnosis is consisting of real-time RT-PCR assay for detection/quantification and type-specific one tube RT-PCR assay for genotyping. Primers and probe were designed based on the consensus sequences in the 5′ UTR and the first 100 bp of the core region of the HCV genome. Both assays can detect genotypes 1–4 as intended. The limit of detection was 112 IU/mL (89–173 IU/mL, 95% CI) for the real-time RT-PCR assay and 295 IU/mL for the genotyping assay (ranging from 130 IU/mL for genotype 1 to 435 IU/mL for genotype 4). Concordance between real-time RT-PCR assay and AMPLICOR HCV v2.0 test in detection of HCV was assessed by testing clinical serum samples. Of 158 samples, 102 were tested positive and 56 were negative by both assays (100% specificity and sensitivity). Quantification sensitivity of the real-time RT-PCR assay was evaluated on 25 samples and the results were compared with AMPLICOR HCV Monitor v2.0 kit. The observed relationship between the results was linear with the correlation coefficient of 0.988. Diagnostics specificity and sensitivity of the genotyping assay was determined by comparing its performance on 102 positive samples with the results obtained by ASO hybridization method. The type-specific RT-PCR assay was able to genotype 99 samples (97%) with 100% specificity while at three samples genotyping failed due to the low viral load under the assay's limit of detection. The overall procedure of HCV diagnosis is performed in a closed system with little risk of contamination and is completed within 6 h. Despite being fast and cost-effective, this approach is straightforward, reproducible and avoids post-PCR enzymatic and hybridization steps while detecting and genotyping HCV with high clinical sensitivity. 1. Zein NN. Clinical significance of hepatitis C virus genotypes. Clinical microbiology reviews. 2000; 13(2): 223–35. 2. Popovski ZT, Dimovski AJ, Simjanovska LJ, Chalovska V, Trajanovski D, Grunevska V, et al. Molecular detection and characterization of hepatitis C virus in the Republic of Macedonia. Maк Meд Пpeглeд 1996; 50: 85–90." @default.
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• W2006104692 date "2013-09-01" @default.
• W2006104692 modified "2023-09-23" @default.
• W2006104692 title "P2: An efficient and low cost method for detection, quantification and genotyping of hepatitis C virus" @default.
• W2006104692 doi "https://doi.org/10.1111/jvh.12166_1" @default.
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